BackgroundMicrobial lipids have drawn increasing attention in recent years as promising raw materials for biodiesel production, and the use of lignocellulosic hydrolysates as carbon sources seems to be a feasible strategy for cost-effective lipid fermentation with oleaginous microorganisms on a large scale. During the hydrolysis of lignocellulosic materials with dilute acid, however, various kinds of inhibitors, especially large amounts of organic acids, will be produced, which substantially decrease the fermentability of lignocellulosic hydrolysates. To overcome the inhibitory effects of organic acids, it is critical to understand their impact on the growth and lipid accumulation of oleaginous microorganisms.ResultsIn our present work, we investigated for the first time the effect of ten representative organic acids in lignocellulosic hydrolysates on the growth and lipid accumulation of oleaginous yeast Trichosporon fermentans cells. In contrast to previous reports, we found that the toxicity of the organic acids to the cells was not directly related to their hydrophobicity. It is worth noting that most organic acids tested were less toxic than aldehydes to the cells, and some could even stimulate the growth and lipid accumulation at a low concentration. Unlike aldehydes, most binary combinations of organic acids exerted no synergistic inhibitory effects on lipid production. The presence of organic acids decelerated the consumption of glucose, whereas it influenced the utilization of xylose in a different and complicated way. In addition, all the organic acids tested, except furoic acid, inhibited the malic activity of T. fermentans. Furthermore, the inhibition of organic acids on cell growth was dependent more on inoculum size, temperature and initial pH than on lipid content.ConclusionsThis work provides some meaningful information about the effect of organic acid in lignocellulosic hydrolysates on the lipid production of oleaginous yeast, which is helpful for optimization of biomass hydrolysis processes, detoxified pretreatment of hydrolysates and lipid production using lignocellulosic materials.
Oleaginous yeast Trichosporon fermentans was proved to be able to use sulphuric acid-treated sugar cane bagasse hydrolysate as substrate to grow and accumulate lipid. Activated charcoal was shown as effective as the more expensive resin Amberlite XAD-4 for removing the inhibitors from the hydrolysate. To further improve the lipid production, response surface methodology (RSM) was used and a 3-level 4-factor Box-Behnken design was adopted to evaluate the effects of C/N ratio, inoculum concentration, initial pH and fermentation time on the cell growth and lipid accumulation of T. fermentans. Under the optimum conditions (C/N ratio 165, inoculum concentration 11%, initial pH 7.6 and fermentation time 9 days), a lipid concentration of 15.8g/L, which is quite close to the predicted value of 15.6g/L, could be achieved after cultivation of T. fermentans at 25°C on the pretreated bagasse hydrolysate and the corresponding lipid coefficient (lipid yield per mass of sugar, %) was 14.2. These represent a 32.8% improvement in the lipid concentration and a 21.4% increase in the lipid coefficient compared with the original values before optimization (11.9g/L and 11.7). This work further demonstrates that T. fermentans is a promising strain for lipid production and thus biodiesel preparation from abundant and inexpensive lignocellulosic materials.
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